• Mycobiology
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• 제39권1호
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• pp.40-44
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• 2011

Aspergillus fumigatus is associated with invasive disease aspergillosis in immunocompromised individuals. The major aim of this study was to investigate the biochemical and immunological responses of male Wistar rats against A. fumigatus experimentally-induced pulmonary fungal infection. Nostril experimental exposure of male Wistar rats to a high dose of A. fumigatus freeze-dried preparation for only 24 hr resulted in a significant increase in levels of catalase, nitric oxide and lipid peroxide in lung homogenates, compared to those of the control animals. However, the oxidative status of the lungs of rats challenged with killed fungus did not change significantly, except for the stimulation in the level of lipid peroxide. IgG level was significantly elevated only in rats that received two low doses of fungus, compared to unexposed animals (p < 0.005). Examining the lung of rats exposed to A. fumigatus revealed no abnormal changes, except for pus in bronchial lumen spaces and per bronchial inflammation. Histologically, large numbers of granuloma cells were evident in the lungs of challenged rats, while no granuloma formation was evident in the lungs of rats exposed to killed fungus.

• Journal of Pharmaceutical Investigation
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• 제21권3호
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• pp.161-170
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• 1991

The storage stabilities of immunoliposomes incorporated with variable amounts of ganglioside $G_{M1}$ were investigated as a function of time. temperature. and composition by observing absorbance of visible light and calcein release. In the column chromatographe, the layer of unsaturated PE(dioleoylphosphatidylethanolamine : DOPE). unable to form stable liposomes at physiological temperature and pH, were formed when palmitoyl-immunoglobulin G(IgG) $(2.5{\times}10^{-4}\;mol/DOPE\;mol)$ added. The incorporation of ganglioside $G_{M1}$ into immunoliposome. enhanced the stabilities of bilayers during the extended period of storage. The turbidities of immunoliposomes coated with ganglioside $G_{M1}$ exhibited the maximum near 20 mol% $G_{M1}/DOPE$ mol. probably because of the disturbance of the bilayer characteristics, i.e., layer transition or reorientation of interaction sites. At low temperature. the higher stability was achieved than at elevated temperatures. After one week of storage. the redispersed liposomal solutions at lower temperatures maintained the original elution patterns in chromatography but broader distribution at elevated temperatures. During the storage, it is suggested the aggregation is the more dominant phenomena for liposomes kept at $5^{\circ}C$ than the fusion. while he fusion is at elevated temperatures.

• KSBB Journal
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• 제25권1호
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• pp.97-102
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• 2010

The enhancement of immunity for atopic dermatitis with application of eicosapentaenoic acid (EPA)-loaded liposome was evaluated on NC/Nga mice. The EPA-loaded liposome was coated with thiol-chitosan. The liposomes were characterized with transmission electron microscopy (TEM), surface zeta potential & particle size analyzer (Zeta-PSA) and differential scanning calorimetry (DSC). The loading efficiency of EPA in the liposome was about 4.7%. The particle size of the EPA-Ioaded liposome was about 230 nm. The values of Immunoglobulin E (IgE), interleukin-4 (IL-4), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) were reduced significantly with application of the EPA-loaded liposome. The interferon-$\gamma$ (IFN-$\gamma$) value was increased with the application effect. It is concluded that EPA loaded liposome have immunity advancing effects in mouse model of atopic dermatitis.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.110-111
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• 2013

FcBP consisting of 13 amino acids specifically binds to Immunoglobulin G Fc domain. Initially, we utilized this peptide for preparation of antibody chip as a PEG composite for enhanced solubility. After then, the peptide conjugate was immobilized on agarose resin, resulting in highly efficient affinity column for antibody purification. The efficiency was comparable to commercial Protein A column. Recently, this peptide was conjugated with cell penetrating peptide (CPP) on a backbone of GFP, affording antibody transducer, which carries antibody into live cells by simple mixing of antibody and the transducer in cell culture media. Antibody transduction into cells was monitored by live cell imaging. More recently, the FcBP was fused to ferritin cage, which consists of 24 ferritin protein molecules. The FcBP-ferritin cage showed greatly increased binding affinity to human IgG. Its binding was analyzed by QCM and SPR analysis. Finally, it was selectively delivered by Herceptin to SKBR3, a breast cancer cell, over MCF10A, non-tumorigenic cells (Fig. 1). Fig. 1. Fluorescent microscopic images of SKBR3 breast cancer cells (A~C) and MCF10A breast cells (D~F) treated with Cy3-trastuzumab/fFcBP-Pf_Fn complexes. Trastuzumab and FcBP-Pf_Fn, which were labeled with Cy3 (Cy3-trastuzumab) and fluorescein (fFcBP-Pf_Fn), respectively, selectively targeted SKBR3 over MCF10A.

• Journal of Microbiology and Biotechnology
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• 제27권4호
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• pp.718-724
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• 2017

The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and half-life. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

• Preventive Nutrition and Food Science
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• 제7권4호
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• pp.373-377
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• 2002

Allergens in processed foods may place persons with food allergies at significant risk when the labels do not Provide sufficient warnings or identification of high-risk ingredients. Because egg proteins are common food allergens, this study was carried out to identify hen's egg albumin (ovalbumin, OVA) in five commercially processed foods containing egg (custayd, cookie and pasta), and chicken meat (sausage and meatball) by immunological methods using commercially produced murine monoclonal immunoglobulin G (M-IgG), immunoblotting and enzyme linked immunosorbent assay (ELISA). Sample buffer with chelating and reducing agents was prepared and used for the preparation of the protein fractions from the foods. Most bands in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profile (5~15% gradient gel) presented at 75 kDa below. OVA (43 kDa) in the sample lanes could not be visually observed on the gel. However, OVA in solutions prepared from custard and cookie could be detected by M-IgG, but were not detected in sausage and pasta. OVA in all samples could be quantitatively determined by the equation obtained from the standard curve by ELISA. Cookie and custard containing egg white and egg, respectively, contained very high concentrations of OVA. OVA in the other products were present in relatively low concentrations, but sufficiently high to pose possible risk of allergy, ELISA is a very sensitive and precise method for the identification and quantification of allergens in food products including allergy-inducible materials.

• 대한본초학회지
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• 제30권3호
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• pp.1-12
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• 2015

Objectives : In this study, we investigated the effects of Agastachis Herba water (AH-W) extract on compound 48/80-induced mast cell degranulation and histamine release in human mast cells and also anti-asthmatic effect of AH-W extract on ovalbumin (OVA)-induced asthma in mice. Methods : Human mast cells, HMC-1 were treated with AH-W extract in the presence or absence of compound 48/80 (C48/80). Mast cell degranulation was observed by microscope, and the histamine release was measured in culture medium by ELISA. For preparation of asthmatic in vivo model, mice were sensitized (0, 7, and 14 days) with OVA and airway challenged (21, 23, 25, 27, and 29 days). AH-W extract at doses of 100 and 300 mg/kg/body weight was orally administered during OVA challenge once per a day. The levels of immunoglobulin (Ig) E, and Th1/Th2 cytokines, IFN-$\gamma$ and IL-4 were measured in the sera of mice by ELISA. The histopathological change of lung tissues was observed by hematoxylin and eosin (H&E) and Periodic Acid Schiff (PAS) staining. Results : The treatment of AH-W extract significantly decreased the mast cell degranulation and histamine release in C48/80-stimulated HMC-1 cells. In addition, The administration of AH-W extract at does of 100 and 300 mg/kg significantly decreased the serum levels of OVA-specific IgE compared with those of OVA control group. In H&E and PAS staining, AH-W extract inhibited OVA-induced airway inflammation, and inflammatory cells infiltration, and also histopathological damages on lung tissues such as bronchiole epithelial desquamation, goblet cells hyperplasia, and mucin releasing. Conclusions : These results indicate that AH-W extract may improve asthmatic symptoms through mast cell stabilization and inhibiting the lung inflammation in bronchial asthma.

• KSBB Journal
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• 제19권1호
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• pp.88-92
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• 2004

장티푸스 협막 다당체와 일본 뇌염 바이러스로 구성되어있는 혼합백신을 제조하여 마우스에서 면역성을 측정하였다. 혼합 항원 백신은 단일 항원을 투여한 경우보다 장티푸스와 일본 뇌염에 대해서 더 높은 IgG 항체 형성을 나타내었다. Aluminium hydroxide를 adjuvant로 첨가한 경우에도 첨가하지 않은 대조군에 비하며 모두 IgG 항체 형성이 증가하는 것으로 나타났다. 일본뇌염의 경우 중화항체 값 측정 시험에서도 혼합 항원백신은 일본뇌염 단독 항원 시에 비교해서 더 많은 중화항체 값을 형성하는 결과를 나타내었다. 본 실험에서 장티푸스 협막 다당체와 일본뇌염 바이러스의 혼합백신은 서로 다른 성분의 항원간에 나타날 수 있는 항체 형성에 대한 masking effect가 발생하지 않으며 오히려 synergic effect를 나타낸다는 사실을 보여주었다. 본 연구에서는 접종시기가 유사한 두 항원의 혼합 백신으로의 사용가능성을 높여주고 최근에 백신 산업에서 종류가 다른 여러 백신들을 혼합하여 다가백신을 제조 생산하는 추세의 산업화에 큰 기대효과를 제공할 수 있으리라 사려된다.

• KSBB Journal
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• 제14권1호
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• pp.82-90
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• 1999

병원성 미생물을 측정할 수 있는 분석시스템을 구성하기 위해 면역학적 성분들을 합성하였고, 이를 이용하여 모델 시스템을 구성함으로써 균 세포 분석원리가 연구되었다. 구성성분을 준비하기 위해 Salmonella thompson에 대한 복합 클론항체를 면역 친화 크로마토그래피를 이용하여 정제하였고, 이렇게 정제된 항체를 Streptavidin과 horseradush peroxdase에 화학결합시켰다. 항체와 Streptavidinfdm은 각각 SMCC와 SPDP에 의해 활성화 되있고 두 물질을 반응시킴으로써 중합체가 합성되었다. 중합체는diaminobiotion 젤과 sephades G-100젤을 이중 층으로 쌓은 칼럼을 이용하여 정제되었다. 항체- HRP 중합체의 합성을 위해, HRP를 $NAIO_4$ 처리에 의해 안정화된 중합체는 Biohel A5M을 이용한size exchusion크로토그래피로 정제되었다. 이렇게 준비된 중합체들과 dot-bloner 그리고 biotim이 고정화된 nitrocellulose membrane($12\mum$ pore size)을 이용하여 모델시스템을 구성하였다. 분석물질(S.Thormpson cells)을 먼적 액상에서 두 중합체와 반응되었고 반응먹을 membrane이 정착된 blotter에 옮긴 후 하부에 진공을 걸어 면역복합체를 biotin-streptavidin 반응에 의해 membrane 표면에 포획하였다.최적조건 하에서 시스템의 균 세포 분석원리를 확인하였으며 측정하한농도는 약 $1{\mu}g/m{\ell}(10^5 {\cdot} 10^6\;cells/m{\ell}$인 것으로 나타났다. 이러한 측정성능의 주요조절인자는 두항체 종합체 농도의 증가는 항원-항체 응집반응을 초래하는 것으로 나타났다.

• 한국식품과학회지
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• 제41권4호
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• pp.405-412
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• 2009

본 연구는 꾸지뽕 나무 열매로부터 75 kDa의 당단백질(꾸지뽕 당단백질)을 추출한 후 꾸지뽕 당단백질의 첨가에 따른 알레르기성 염증 인자인 histamine 유리 억제능력 및 COX-2의 활성 억제 효과를 평가하였다. RBL-2H3세포를 22시간 동안 IgE로 감작시킨 후, HSA를 처리하여 histamine의 유리양을 측정한 결과 꾸지뽕 당단백질을 처리한 농도가 증가함에 따라 histamine의 유리와 COX-2의 활성 억제율은 증가하였다. 또한 꾸지뽕 당단백질의 처리는 HSA에 의해 유도된 세포내 ROS 생성량을 농도에 의존적으로 억제하였다. 한편 꾸지뽕 당단백질을 농도별로 처리하여 세포내 단백질을 추출하여 western blot을 실시한 결과 100 ${\mu}g$/mL 농도의 꾸지뽕 당단백질을 처리한 그룹에서 ERK1/2, AP-1과 COX-2의 활성 수준은 현저히 억제 되었다(p<0.05). 따라서 이러 한 결과에 미루어볼 때, 꾸지뽕 당단백질은 세포내 해독효소의 활성을 증가시킴으로써 ROS 수준을 감소시켰으므로 꾸지뽕 당단백질의 역할이 다른 천연물 유래의 당단백질과 마찬가지로 특이적인 항산화 능력을 지니고 있음을 나타내며 histamine의 유리 억제와 COX-2의 활성이 억제되었을 것으로 생각된다. 이는 꾸지뽕 당단백질이 항 알레르기 효능을 갖는 물질로써 알레르기성 비염, 아토피 등과 같은 알레르기 관련 질환의 예방 및 치료제로 사용될 수 있을 것 사료된다.